Structural mechanism of CRL4-instructed STAT2 degradation via a novel cytomegaloviral DCAF receptor

EMBO J. 2023 Mar 1;42(5):e112351. doi: 10.15252/embj.2022112351. Epub 2023 Feb 10.

Abstract

Human cytomegalovirus (CMV) is a ubiquitously distributed pathogen whose rodent counterparts such as mouse and rat CMV serve as common infection models. Here, we conducted global proteome profiling of rat CMV-infected cells and uncovered a pronounced loss of the transcription factor STAT2, which is crucial for antiviral interferon signalling. Via deletion mutagenesis, we found that the viral protein E27 is required for CMV-induced STAT2 depletion. Cellular and in vitro analyses showed that E27 exploits host-cell Cullin4-RING ubiquitin ligase (CRL4) complexes to induce poly-ubiquitylation and proteasomal degradation of STAT2. Cryo-electron microscopy revealed how E27 mimics molecular surface properties of cellular CRL4 substrate receptors called DCAFs (DDB1- and Cullin4-associated factors), thereby displacing them from the catalytic core of CRL4. Moreover, structural analyses showed that E27 recruits STAT2 through a bipartite binding interface, which partially overlaps with the IRF9 binding site. Structure-based mutations in M27, the murine CMV homologue of E27, impair the interferon-suppressing capacity and virus replication in mouse models, supporting the conserved importance of DCAF mimicry for CMV immune evasion.

Keywords: cullin-RING ubiquitin ligases; cytomegalovirus; interferon; ubiquitin-proteasome system; viral DCAF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cryoelectron Microscopy
  • Cytomegalovirus Infections* / genetics
  • Humans
  • Interferon-Stimulated Gene Factor 3, gamma Subunit / metabolism
  • Interferons / metabolism
  • Mice
  • Muromegalovirus*
  • Rats
  • Receptors, Interleukin-17 / metabolism
  • STAT2 Transcription Factor / genetics
  • STAT2 Transcription Factor / metabolism
  • Ubiquitin-Protein Ligases / metabolism

Substances

  • 2,4-bis(N,N'-di(carboxymethyl)aminomethyl)fluorescein
  • Interferon-Stimulated Gene Factor 3, gamma Subunit
  • Interferons
  • IRF9 protein, rat
  • STAT2 protein, human
  • STAT2 Transcription Factor
  • Ubiquitin-Protein Ligases
  • Receptors, Interleukin-17

Associated data

  • PDB/7zn7
  • PDB/7znn